Figure 3.

(a) Images obtained from the immunocytochemistry experiments in cultured adhered cells (top images) and in cells released into solution inside the device (bottom images). ICC experiments were performed using MCF-7, MDA-MB-435, SKBR3 and PBMCs. Each cell line was tested individually in glass slides (top); additionally, a cell mixture of the different cell lines was spiked in healthy whole blood samples and processed in the device (bottom). MCF-7 cells stained strongly for CK, with cytoplasm localization; SKBR3 cell membrane stained positively for HER2; most PBMCs stained for CD45. (b) For further image analysis, CTC classification was established according to the combination of the presence and absence of the selected biomarkers, and CD45 is used as CTC exclusion criteria.